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目的：建立适合本实验室条件的非同位素ＰＣＲ－ＳＳＣＰ－ＲＦＬＰ基因突变分析技术。方法：应用银染ＰＣＲ－ＳＳＣＰ技术检测石蜡包埋复发肝细胞癌（Ｈｅｐａｔｏｃｅｌｕｌａｒｃａｒｃｉｎｏｍａ，ＨＣＣ）组织中Ｐ５３基因第七外显子突变。采用系列步骤克服非特异扩增；分析影响ＳＳＣＰ诸参数的优化；并与ＲＦＬＰ结果相印证。结果：检测复发ＨＣＣ标本２２例，ＳＳＣＰ阳性率７７．２７％（１７／２２），２４９位点突变率７２．７２％（１６／２２），ＲＦＬＰ与ＳＳＣＰ分析结果符合率１００％；另有２例发生２４９位点以外的突变。结论：该实验流程可靠、高效，适合大量标本的基因突变筛查
Objective: To establish non-isotopic PCR-SSCP-RFLP gene mutation analysis techniques suitable for the conditions of our laboratory. METHODS: The mutation of exon 7 of P53 gene in paraffin-embedded and recurrent hepatocellular carcinoma (HCC) tissues was detected by silver staining PCR-SSCP. A series of steps were used to overcome non-specific amplification; the analysis affected the optimization of SSCP parameters; and corroborated with RFLP results. Results: Twenty-two patients with recurrent HCC were detected. The positive rate of SSCP was 77.27% (17/22). The mutation rate of 249 sites was 72.72% (16/22). The coincidence rate of RFLP and SSCP analysis was 100%; There were mutations outside the 249 site. Conclusion: The experimental procedure is reliable, efficient, and suitable for genetic mutation screening of a large number of specimens